HA751726

GLS2 Recombinant Rabbit Monoclonal Antibody [PSH19-71]

cat.: HA751726

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, IP
Clonality:	Monoclonal
Clone number:	PSH19-71

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4).
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 66 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within human GLS2 aa 301-350.
Positive control:	HepG2 cell lysate, MCF7 cell lysate, NCI-H1299 cell lysate, T-47D cell lysate, Mouse liver tissue lysate, Mouse pancreas tissue lysate, Rat liver tissue lysate, Rat pancreas tissue lysate, HepG2, human liver tissue, mouse liver tissue, rat liver tissue.
Subcellular location:	Mitochondrion.
Recommended Dilutions: WB IF-Cell IHC-P IP	1:5,000-1:20,000 1:50 1:200 1-2μg/sample
Uniprot #:	SwissProt: Q9UI32 \| Q571F8 \| P28492
Alternative names:	breast cell glutaminase GA GLS Gls2 GLSL_HUMAN glutaminase 2 (liver mitochondrial) glutaminase 2 glutaminase GA glutaminase I Glutaminase liver isoform Glutaminase liver isoform, mitochondrial hLGA L glutaminase L glutamine amidohydrolase L-glutaminase L-glutamine amidohydrolase LGA mitochondrial phosphate activated glutaminase phosphate-dependent glutaminase

Images

Fig1: Western blot analysis of GLS2 on different lysates with Rabbit anti-GLS2 antibody (HA751726) at 1/5,000 dilution.

Lane 1: HepG2 cell lysate
Lane 2: MCF7 cell lysate (low expression)
Lane 3: NCI-H1299 cell lysate (low expression)
Lane 4: T-47D cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 66 kDa
Observed band size: 60 kDa

Exposure time: 30 seconds; ECL: K1801;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751726) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of GLS2 on different lysates with Rabbit anti-GLS2 antibody (HA751726) at 1/5,000 dilution.

Lane 1: Mouse liver tissue lysate
Lane 2: Mouse kidney tissue lysate (negative)
Lane 3: Mouse pancreas tissue lysate
Lane 4: Rat liver tissue lysate
Lane 5: Rat kidney tissue lysate (negative)
Lane 6: Rat pancreas tissue lysate

Lysates/proteins at 40 µg/Lane.

Predicted band size: 66 kDa
Observed band size: 60 kDa

Exposure time: 30 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751726) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunocytochemistry analysis of HepG2 cells labeling GLS2 with Rabbit anti-GLS2 antibody (HA751726) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-GLS2 antibody (HA751726) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.
	Fig4: Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-GLS2 antibody (HA751726) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA751726) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig5: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-GLS2 antibody (HA751726) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA751726) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Fig6: Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-GLS2 antibody (HA751726) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA751726) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Fig7: GLS2 was immunoprecipitated from 0.2 mg HepG2 cell lysate with HA751726 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751726 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HepG2 cell lysate (input)
Lane 2: HA751726 IP in HepG2 cell lysate
Lane 3: Rabbit IgG instead of HA751726 in HepG2 cell lysate

Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 1 minute 34 seconds; ECL: K1801

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.